




































In ternationa l
Scholars
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African Journal of Pig Farming ISSN 2375-0731 Vol. 6 (12), pp. 001-009, December, 2018. Available online at 
www.internationalscholarsjournals.org © International Scholars Journals 

 

Author(s) retain the copyright of this article. 

 
 

Full Length Research Paper 

 

Prevalence, haemolysis and antibiograms of 
Campylobacters isolated from pigs from three farm 

settlements in Venda region, Limpopo province, 
South Africa 

 
P. O. Uaboi-Egbenni1*, P. O. Bessong2, A. Samie3 and C. L. Obi4 

 
1
Department of Microbiology, University of Venda, P.M.B. 5050, Thohoyandou, Limpopo Province, South Africa. 

2
Department of Microbiology, University of Venda, P.M.B. 5050, Thohoyandou, Limpopo Province, South Africa. 

3
Department of Microbiology, University of Venda, P.M.B. 5050, Thohoyandou, Limpopo province, South Africa. 

4
Academic Affairs and Research, Walter Sisulu University, Mthatha, Eastern Cape, South Africa. 

 
Accepted 14 May, 2018 

 
We investigated the prevalence, haemolytic activities and antibiotic susceptibility profiles of Campylobacter 
species isolated from pigs in three farms in the Venda region, Limpopo province, South Africa. During the period of 
investigation, which spanned over one year, 450 faeces samples from pigs comprising 150 each from the three 
farms were collected and appropriately transported to the base laboratory at the Department of Microbiology, 
University of Venda for analysis. In total, the prevalence rate of Campylobacter isolates from all three farms was 
30.2%, whereas, the rates of 25 - 26% were recorded for farms X, Y and Z, respectively. Out of the isolated 
Campylobacter species 10 (12.5%) were identified as Campylobacter jejuni and 70(87.5%) as Campylobacter coli. Of 
the Campylobacter isolates from the farms, 23, 22 and 35 were β-haemolytic from farms X, Y and Z, respectively. 
Susceptibility of the isolates to 12 antibiotics was determined by disc diffusion technique. The overall rate of 
resistance determined in this study to most of the antibiotics was 84.2%. Most Campylobacter isolates from farm X 
(< 50%) were resistant to ciprofloxacin, cefexime, and erythromycin, while > 50% were resistant to tetracycline, 
(54.5%) to vancomycin, (100%) to ampicillin and methicillin. The rates of resistance to these antibiotics were not 
significantly different from farms X and Y (p < 0.005). However, most isolates from farm Y were more resistant to 
ciprofloxacin (57.1%) than isolates from farm X (36.4%) and Z (40.7%). Resistance rates to tetracycline by C. coli 
isolated from farm Y (48%), farm Z (45.5%) and farm X (44.4%) were noted. Similar trend was observed for C. coli 
from farm Y (40%), farm Z (31.8%) and farm X (22.2%) for ciprofloxacin. All C. coli from farm Z showed higher level 
of resistance to ampicillin (100%), Y (64%) and X (55.6%). The C. jejuni isolates though few were markedly resistant 
to fluoroquinolone and the macrolide as well as aminoglycoside antibiotics. Most of the isolates were resistant to 
at least seven (7) of the antibiotics tested. These observations indicate high levels of resistance to antibiotics 
commonly used in the swine industry (erythromycin and tetracycline) among the C. coli and C. jejuni isolates from 
pigs. Erythromycin and ciprofloxacin are drugs for treatment of human campylobacteriosis. The high prevalence 
rate of resistance to these drugs among C. coli and C. jejuni isolates from pig faeces is of public health 
significance. 

 

Key word: Campylobacter, campylobacteriosis, haemolytic, susceptibility, resistance, profiles. 

 
INTRODUCTION 

 
Campylobacter, mainly Campylobacter jejuni and  
 

 
*Corresponding author. E-mail: uaboiegbenni@yahoo.com. Tel: 
+27 15 962 8186. 

 
 
 
 

 
Campylobacter coli, are recognized as major causes of 
acute bacterial food-borne gastroenteritis (World Health 
Organization, 2002). The incidence of human campylo-
bacteriosis in the European Union has increased over the 
past years and recently exceeded that of Salmonella in 



 
 
 

 

many countries. Although campylobacteriosis is usually a 
self-limiting diarrheal disease, severe complications such 
as septicemia, reactive arthritis, and Guillain-Barre 
syndrome sometimes occur (Humphrey et al., 2007).  

C. jejuni colonize the intestinal tracts of a large number 
of mammals and birds. Broilers are often carriers of C. 
jejuni. In Thailand, Campylobacter species were isolated 
from 12% of various food samples including pork, chicken 
and vegetables in Bangkok (Rasrinual et al., 1988; 
Young, 2005), 40% poultry ceca in India (Das et al., 
1996), and 68-100% of poultry samples from retail 
markets in Taiwan (Shih, 2000).  

Reports on Campylobacter in pigs revealed higher 
proportions of C. coli than C. jejuni, whether they have 
enteritis or not (Harvey et al., 1999; Steinhauserova et al., 
2001). In the U.S., Campylobacter species were isolated 
from 76% gilts, 100% of pregnant sows, 57.8% of 
newborn piglets, and 100% of weaning pigs (Young et al., 
2000). In the Netherlands, fifty-percent of piglets were 
infected with the same serotypes at seven days of age 
(Weijtens et al., 1997). The average number of Campylo-
bacter colonizing the gut decreased toward the end of the 
rearing period (Weijtens et al., 1999). A study in Belgium 
reported the prevalence of Campylobacter sp. on pig 
carcasses at slaughterhouses to be 2% (Korsak et al., 
1998), and Campylobacter was found in 1.3% of samples 
from pork from a retail market in U.S. (Duffy et al., 2001). 
Although the poultry industry has often be considered 
primarily responsible for human campylobacteriosis, the 
pork industry has equally been identified as a potential 
source of human infection (Allos, 2001; Berndtson et al., 
1996). Uaboi-Egbenni (2008) also documented the 
prevalence of campylobacters in dogs and guinea-fowl in 
Nigeria.  

The gastrointestinal tracts of other food animal species 
have also been shown to be frequently colonized with 
campylobacters, particularly, C. jejuni and C. coli 
(Minihan et al., 2004). Reported rates of Campylobacter 
carriage in food animals have varied widely between 
studies (Busato et al., 1999). The high prevalence of 
campylobacters in pigs has been reported previously in 
numerous studies and dressed pig carcasses have been 
shown to be more frequently contaminated than either 
beef or sheep (Nesbakken et al., 2003). This is most 
likely attributable to the fact that pig carcasses undergo a 
communal scalding process combined with the fact that 
the skin remains on the carcass following all of the 
dressing procedures (Moore et al., 2005).  

Thermotolerant campylobacters (C. jejuni/C. coli) 
constitute the most frequent cause of intestinal infections 
worldwide. The clinical spectrum of Campylobacter enteritis 

ranges from a watery, nonbloody, noninflammatory diarrhea 

to a severe inflammatory diarrhoea with abdominal pain 
and fever. Disease is less severe in developing countries 
than in developed countries. The disease is characterized 
by bloody stool, fever, and abdominal pain that is often 
more severe than that observed in Shigella and Salmonella 

 
 
 
 

 

infections. In developing countries the features reported 
are watery stool, fever, abdominal pain, vomiting, dehy-
dration, asthenia, anorexia and presence of faecal 
leukocyte; patients are also often underweight and 
malnourished (Rao et al., 2001; Coker et al., 2002). Stools 

remain positive for several weeks.  
Treatment appears to be beneficial if it is administered 

early enough in the course of the disease (Salazar-Lindo 
et al., 1986). The recommended drugs are erythromycin, 
or amoxicillin or a fluoroquinolone (ciprofloxacin, nor-
floxacin) or tetracycline, provided the bacterium has not 
acquired a resistance. However, the emergence of 
antibiotic resistant strains has further opened a new 
dimension as to how to combat the disease together with 
the emerging resistant strains. In 1988, the emergence of 
fluoroquinolone resistance in Campylobacter strains was 
first reported in Spain, and since then the emergence of 
fluoroquinolone resistance has been identified in many 
countries, including Finland, the Netherlands, England, 
and Canada (Berndston et al., 1996), but not in South 
Africa. In Spain, 100% of Campylobacter strains isolated 
from broilers and pigs were found to be resistant to 
fluoroquinolones (Berndtson et al., 1996).  

There is a dearth of information and research on the 
prevalence, epidemiology and antibiograms of campylo-
bacters in pigs in South Africa in spite of the reports in 
Europe and other parts of the world on pigs as potential 
sources of infections for humans. This study therefore 
ascertained the prevalence, antibiotic susceptibility 
profiles and haemolytic activities of campylobacters 
isolated from pigs in three farms in the Venda region, 
Limpopo province of South Africa in order to provide 
updated information and data on the research subject. 

 

MATERIALS AND METHODS 
 
Collection of faeces 
 
A total of four hundred and fifty (450) freshly voided faeces 
comprising one hundred and fifty from each of the three farms were 
collected at random from pigs in sterile 50 ml plastic containers and 
were adequately kept in coolers packed with crushed ice, stored at 
4°C and immediately transported to the laboratory for analysis. The 
faeces were immediately processed. About 2 gm of the samples 
was transferred to 6 ml of sterile phosphate-buffered saline and left 
to emulsify at room temperature for 10 - 20 min to release the 
bacteria. The suspension and bacteria were used directly for the 
identification of Campylobacter. The three (3) sampling sites are 
separated from each other by about 200 km but are located in the 
same province. The three farms were code named X, Y, and Z for 
ethical reasons. 
 
 
Isolation and identification of Campylobacter sp. by 
conventional culture methods 
 
Ten microlitres of the faecal suspension was spread on the surface 
of a charcoal cefoperazone deoxycholate agar plates (CM 739 
[Oxoid] with cefoperazone supplement SR 155E). The plates were 
incubated under microaerophilic conditions employing the Cam-
pygen gas generating kit (Oxoid CM025) at 42°C for 48 h. 



 
 
 

 

Colonies suspected to be Campylobacter were further purified on 
blood agar plates (Blood Agar Base No.2 (Oxoid) supplemented 
with 5% sterile laked horse blood). All the isolates were characterized by 
their catalase, oxidase reactions, hydrogen sulphide production, indoxyl 
acetate, hippurate and susceptibility to nalidixic acid by standard 
procedures (On and Holmes, 1991, 1992). The resulting isolates were 
subsequently stored at -80°C in brain heart infusion broth with 15% 
glycerol until further investigation. 
 

 
Confirmation of presumptive positive Campylobacter strains 

 
Presumptive identification of Campylobacter isolates was done 
using the Dryspot Campylobacter test kit (Oxoid Basingstoke, 
Hampshire England). The test is specific for pathogenic Campylo-
bacter strains belonging to C. jejuni, C. coli, Campylobacter 
upsaliensis, and Campylobacter lari. However, the other 
Campylobacter species are not included in this identification 
scheme. The manufacturer’s instructions were strictly adhered to in 
this procedure. Briefly, 48 h cultures of suspected Campylobacter 
showing the usual cultural characteristics were selected. The 
reagents were removed from the refrigerator and allowed to attain 
room temperature according to the manufacturer’s instructions. The 
extraction tubes were placed in test tube rack. Thereafter, one drop 
of extraction reagent 1 was added into the extraction tube. 
Sufficient growth of the suspected Campylobacter culture was 
removed to fill the inner diameter of a sterile loop. These cells were 
thoroughly suspended in a drop of reagent 1. The loop was allowed 
to stay in this reagent for 3 min. Two drops of extraction reagent 2 
was added to the previous mixture and mixed thoroughly using the 
retained loop. With the aid of a paddle pastette provided with the kit, 
1 drop (50 µl) of the neutralized extract was placed onto the test 
circle and a drop onto the control circle. With the aid of the flat end 
of the paddle pastette, the extract was mixed into the dry control 
reagent spot until completely suspended to cover the reaction area. 

The same pastette was used to repeat this procedure for the test reagent. 
The test cards were rocked for about 3 min. This process was repeated 
for all suspected Campylobacter isolates. Agglutination under normal 
lighting condition indicated that the organism was Campylobacter. 

 
The presumptive positive campylobacters were further subjected 

to Mast diagnostic Campylobacter kits consisting of urease, indoxyl 
acetate and hippurate test solution. Briefly, 24 h cultures of the 
Campylobacter were inoculated into the urease and hippurate test 
solutions according to the manufacturer’s instructions. These were 
then incubated for 4 h for colour development. For urease, 
development of pink colour was indicative of urease enzyme 
production (C. lari), while development of pink colour in hippurate 
solution indicated production of hippuricase enzyme (C. jejuni). In 
the case of indoxyl acetate solution, change of colour from 
colourless to blue/purple was indicative of the presence of C. jejuni 
or C. coli. The indoxyl acetate strips were impregnated with wet 
cultures and allowed to stay for 3 - 5 min. Development of purple 
colour within this period was indicative of positive reaction (C. jejuni 
and C. coli) (Popovic-Uroic et al., 1990; On and Holmes, 1992; 
Elviss et al., 2008). 
 

 
Blood haemolysis test 

 
To ascertain pathogenicity of isolates, the identified Campylobacter 
sp. were subjected to haemolytic test according to the procedure of 
Samie et al. (2007). Briefly, a 24 h broth culture of Campylobacter 
sp. were cultured by spread plate method on Columbia agar 
supplemented with defibrinated sheep red blood cells. Plates were 
incubated at 37 ± 2°C for 24 h. Thereafter, plates were observed for 
complete, partial and no haemolysis. 

 
 

  
 
 

 
Antimicrobial agents 

 
The antibiotics tested in this study were: Trimethroprim (2.5 µg), 
nalidixic acid (30 µg), ciprofloxacin (5 µg), gentamycin (10 µg), 
tetracycline (30 µg), ampicillin (10 µg), erythromycin (15 µg), 
streptomycin (10 µg), methicillin (µg), cefexime (30 µg), imipenem 
(µg), kanamycin (30 µg) and vancomycin (30 µg) (Oxoid, Unipath 
Ltd, Basingstoke, England). 

 

Antimicrobial sensitivity testing 
 
The method of Gaudreau and Gilbert (1997) was used. Briefly, the 
confirmed Campylobacter isolates were inoculated into plates of 
Mueller-Hinton agar supplemented with 5% sheep red blood cells in 
plates carrying a maximum of five (5) antimicrobial discs. All plates 
were incubated at 35°C under a microaerophilic atmosphere obtained with a 
gas generator envelope (Oxoid, Unipath Ltd, Basingstoke, England) for 
24 h. The resulting zone diameters were measured with a graduated 
metre rule. 

 

Preparation of bacterial genome 
 
Genomic DNA was obtained by the whole-cell lysate method as 
described by Marshall et al. (1999). Briefly, cells from a 24 - 48 h 
culture grown on Columbia blood agar were resuspended in sterile 
distilled water to an optical density of 2.5 at 540 nm. The 
suspensions were boiled 100°C for 20 min in Eppendorf tube. The 
resulting templates were either used immediately for polymerase 
chain reaction (PCR) or were kept at 4°C for up to 1 month. 

 

PCR identification of Campylobacter strains 
 
In order to ascertain if the dryspot positive Campylobacter isolates 
were genuinely campylobacters, they were subjected to PCR 
identification using the general primers for the identification of 
Campylobacter. These primers are also specific for other members 
of the campylobacteriaceae (Helicobacter and Arcobacter). 
However, Arcobacter and Helicobacter sp. show negative reaction 
to the Campylobacter dryspot kit. Hence, any amplification of the 
primer sequences at the 1,004-bp fragment within the coding region 
of 16S rRNA confirmed such isolates as Campylobacter sp. and not 
Helicobacter or Arcobacter sp. The PCR-restriction fragment length 
polymorphism (RFLP) method used in this study was as previously 
described by Marshall et al. (1999). Briefly, amplification was done 
in 50-µl reaction volume containing 5 µl of whole-cell lysate, 1 µl 

each primer, 10x buffer (Roche), 1.5 mM MgCl2, 200 µM each 

deoxynucleotide (Roche) and 5U Taq DNA polymerase (Roche). 
The PCR amplification was performed with a – thermocycler (ESCO 
Swift Mini Thermal Cycler Version 1.0, ESCO Technologies, 
Philadelphia U.SA). The samples were subjected to an initial dena-
turation for 2 min at 95°C, followed by 30 amplification cycles, each 
consisting of 94°C for 30 s, 52°C for 30 s, and 72°C for 90 s. A final 
primer extension at 72°C for 10 min was included. Oligonucleotides 
primers employed in this study were CAH16S 1a (5’ – AAT ACA 
TCA AAG TCG AAC GA – 3’) and CAH16S 1b (TTA ACC CAA CAT 
CTG ACG AC – 3’), respectively. The oligonucleotides used in this study 
were synthesized by Roche Applied Science (Manheim, Germany). 
 

 

RESULTS 
 
Cultural studies 
 
A total  of  four  hundred  and  fifty  (450)  freshly  voided 



      
 
 
 
 

 
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Campylobacters isolated from pig faeces in farms X, Y and Z 
 

 
Fig.1 : Shows the number of C. jejuni , C. coli, C. lari and their percentages isolated from three pig farms in South Africa 

 
 
 

100                                               
 

                  89.7                      
 

90                                               
 

                                              
 

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Farm Y 

 
 

                            

   

  

                             
 

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Fig. 2: Results of haemolysis of sheep red blood cells by Campylobacter isolates              
  

 

 

faecal samples were processed for the detection of 
campylobacters over the course of the study. Of the 138 
presumptive isolates from pigs on mCCDA from the 
farms, 25.6% (115 out of 450) were confirmed as 
campylobacters. Figures 1 and 2 show the percentage 
incidence of Campylobacter strains from pig faeces in the 
three (3) farms. From, there was an equal percentage of 
occurrence of campylobacters in farms X and Y with a 
value of 25.3%. However, there was a higher incidence of 
campylobacters among pigs in farm Z (26%) than in 
farms X and Y. It must be noted here that some strains 

 
 

 

that could not be resuscitated did not form part of the 
analysis and subsequent studies. With the aid of the 
Campylobacter dryspot and Mast diagnostic kits, 2(5.3%) 
of the isolates from farm X were identified as C. jejuni, 
while 21 (55.3%) were identified as C coli. In farm Y, 3 
(7.9%) were C. jejuni, while 19 (50%) were C. coli. In 
farm Z, 5 (12.3%) were C. jejuni, while 30 (76.9%) were 
C. coli. Overall, the prevalence of C. jejuni among pigs in 
the three farms was 10 (12.5%), while it was 70 (87.5%) 
for C. coli (Figure 2). From this finding, there was a 
preponderance of C. coli compared with C. jejuni in all 



   
 
 
 
 
 

 

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100%            

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31.8 
 

45.
5 

           

50 

        

50 

        
 

90% 

                                
 

                                  
 

        

100 

  

100 

 

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0 

 

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    FARM Z R  

                          
 

                             
 

                             FARM Z S  

60% 
  

40 
  

48 
  

48 
    

64 

 

64 

  

52 
  

40 
     

60 

        
 

                 

72 

          

FARM Y R 

 

50%            96              
10
0  

10
0     

 

40% 
  

60 
  

52 
  

52 
         

48 
  

60 
     

40 
        FARM Y S 

 

          
36 

 
36 

      
28 

          
FARM X R  

                                
 

                              

0  

0       

30%   22.2  

44.
4 

  

33.3 
 11.1               11.1       

 

          

55.6 

 

77.8 

 

55.
6 

              

FARM X S 

 

20% 

        

88.9 

10
0 

     

10
0 

  

88.
9 

10
0 

 

10
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77.8 

                  
 

      

66.7 

                  

                       
 

10% 

   

55.
5 

                     
 

                            
 

                                
 

            

44.4 
    44.

4 
               

 

0% 
             

0 
  

22.2 
   

0 
    

0 
       

 

                                
 

                                  
 

   CIP  TE CFM  IPM VA AMP  CN  K MET  E W NA     
   

Antimidrobials to which C.coli isolates were exposed 
 

 
Fig. 3: Percentage susceptibility profile of Campylobacter coli exposed to 12 antibiotics (Key: S = Susceptibility; R = 
Resistance; CIP= Ciprofloxacin; TE= Tetracycline; CFM = Cefexime; IPM = Imipenem; VA= Vancomycin; AMP = 
Ampicillin; CN= Gentamycin; K= Kanamycin; MET= Methicillin; E=Erythromycin; W= Trimethoprim; NA= Nalidixic acid). 

 

farms. 
 

 

Blood haemolysis 

 

Of the 115 Campylobacter strains from the pigs tested for 
sheep blood haemolysis, 23 were β-haemolytic, 10 were 
α-haemolytic and 5 non-haemolytic in farm X, 22 were β-
haemolytic, 16 were α-haemolytic for farm Y, 35 were β-
haemolytic and 4 were α-haemolytic for farm Z (Figure 3). 
 

 

Antimicrobial resistance 

 

Sixty-six isolates were tested for their antibiotic sus-
ceptibility profiles. Most Campylobacter isolates from farm 
X (< 50%) were resistant to ciprofloxacin, cefexime, and 
erythromycin, while > 50% were resistant to tetra-cycline 
(54.5%), vancomycin (100%), ampicillin (54.5%), 
kanamycin (63.6%), and methicillin (100%). In farm Y, ≥ 
50% were resistant to ciprofloxacin (57.1%), tetracycline 
(50%), cefexime (53.6%), vancomycin (67.9%), gentamycin 
(53.6%), methicillin (75%), and erythromycin (53.6%), 
except imipenem and kanamycin which were 10.7 and 
46.4% resistant, respectively. In farm Z, > 50% but < 
100% were resistant to tetracycline, gentamycin and 
erythromycin, while 40.7% were resistant to ciprofloxacin. 
Approximately 100% of Campylobacter species in this 
farm were resistant to cefexime, vancomycin, kanamycin, 
and methicillin. Resistance to most of the antibiotics tested 

was more pronounced among isolates from farm Z. 
Majority of the isolates from farm Y were more resistant 
to ciprofloxacin (57.1%) than isolates from farm X 

 

 

(36.4%) and farm Z (40.7%) (Figure 4 and 5). There were 
no significant differences in resistance profiles amongst 
Campylobacter isolates in the farms (p < 0.005). Study of 
individual species resistance showed that in farm X, all C. 
jejuni isolates were resistant to Ciprofloxacin, tetracycline, 
cefexime, imipenem, vancomycin, gentamycin, kanamycin 

and methicillin. Of the three C. jejuni isolates from farm Y, 
≤ 100% were resistant to all the antimicrobials except 
tetracycline (33.3%) and nalidixic acid in which all were 
susceptible. In farm Z, ≤ 100% were resistant to 10 of the 
antimicrobials except ciprofloxacin to which 80% were 
resistant and nalidixic acid to which 100% were 
susceptible. For C. coli, in farm X < 50% were resistant to 
ciprofloxacin, tetracycline, cefexime, imipenem and 
erythromycin except vancomycin, ampicillin, gentamycin, 
kanamycin, methicillin and trimethoprim to which ≤ 100% 
were resistant. In farm Y, < 50% of C. coli were resistant 
to ciprofloxacin, tetracycline, cefexime, imipenem and 
kanamycin, while ≤ 100% were resistant to vancomycin, 
ampicillin, gentamycin, methicillin, erythromycin and 
trimethoprim. In farm Z, ≤ 50% were resistant to 
ciprofloxacin, tetracycline, gentamycin and erythromycin, 
while all C. coli isolates (100%) were resistant to cefexime, 
vancomycin, ampicillin, kanamycin, methicillin and 
trimethoprim. Analysis of the zone diameters was done 
using CLSI/NCCLS (2002) for enterobacteriaceae.  

The PCR micrographs of the DNA from Campylobacter 
strains from sheep are as indicated in. The purified DNA 
from the Campylobacter strains amplified at the 1004 bp, 
which is the specific region for the conserved 16S rRNA 
for members of the genus Campylobacter. The bands 
formed were confirmed as those for Campylobacter. 
Specific identification by the Mast diagnostic kits 



   
 
 
 

 
100%        0                     

 

                    20         
 

90%  

80 
                          

 

   100  100  100  100  100  100  100  100    100  100     
 

80%                    80        FARM Z R 
 

70%  20  0  0    0  0  0  0  0    0  0    FARM Z S 
 

60%  
66.7 

 33.3    
66.7 

   
66.7 

 
66.7 

             
FARM Y R  

50% 

    

100 

  

100 

   

100 

 

100 

 

100 

 

100 

 

100 

 

  

 

  

66.7 

           
 

               
 

                   FARM Y S  

40% 
 

33.3 
    

33.3 
   

33.3 
 

33.3 
             

 

    0   0    0  0  0  0  0    FARM X R 
 

30%            

50 
       

50 
        

 

20% 
 

100 
 

100 
 

100 
 

100 
 

100 
  

100 
 

100 
 

100 
  

100 
 

100 
   

FARM X S 
 

               
 

                 
 

                  
 

10% 
      

50 
    

50 
       

                          
 

                           
 

0%  0  0  0  0  0    0  0  0    0  0     
 

                  
 

  CIP  TE  CFM IPM  VA  AMP  CN  K  MET  E  W  NA    
   

Antimicrobials to which Campylobacter jejuni were exposed 
 

 
Fig.4: Percentage susceptibility profile of Campylobacter coli exposed to 12 antibiotics (Key: S = Susceptibility; R = 
Resistance; CIP= Ciprofloxacin; TE= Tetracycline; CFM = Cefexime; IPM = Imipenem; VA= Vancomycin; AMP = 
Ampicillin; CN= Gentamycin; K= Kanamycin; MET= Methicillin; E=Erythromycin; W= Trimethoprim; NA= Nalidixic acid).  

 
 
 
 
 
 
 
 

 

1004bp 
 

1004bp 
 
 
 
 
 
 

 

Fig.5: PCR products of amplified DNA from pig Campylobacter isolates aligning at the 1004bp of a 1.9kb ladder (a) pig and 
(b). (a) Lane 1= 1.9kb ladder; lane 2,3,4,5,6,7, amplified bands of DNA from pig Campylobacter strains; (b) lane 1= 1.9kb 
DNA ladder; lanes 2,3,4,5,6,7 are amplified bands of DNA from pig (farm Y) Campylobacter strains. 

 

 

differentiated the isolates into C. jejuni and C. coli. 
 

 

DISCUSSION 

 

Elucidating the shedding patterns and prevalence of 
Campylobacter strains in the faeces of farm animals is a 
prerequisite for effective healthcare strategy against 
zoonotic infections. In the current study, the prevalence of 
campylobacters from pigs and their antimicrobial 
susceptibility pattern was determined. Of the 450 fresh 
faecal samples processed for all farms, 84.6% were 
identified as campylobacters. There was an equal pre-
valence (25.3%) among pigs’ faeces in farm X and Y, but 
a slightly higher predominance in faeces from farm Z. In 
an epidemiological study carried out on 95 pigs 
originating from eight farms, Weijtens et al. (1993) found 

 
 

 

11% of the stomach samples of pigs to be positive for 
Campylobacter, whereas, 85% of faeces were positive. A 
prevalence of campylobacters in pigs ranging from 63 - 
100% has been reported in other studies (Weijtens et al., 
1993; Saenz et al., 2000; Young et al., 2000; Pezzotti et 
al., 2003; Payot et al., 2004). In our findings, none of the 
farms was free from Campylobacter. There were no 
significant differences in the prevalence of Campylo-
bacter sp. in the three farms (X, Y, Z) with the average 
prevalence value at 25.3, 25.3 and 26% respectively (p < 
0.005). Weijtens et al. (1993, 1999) reported high counts 
of shedding of Campylobacter genotypes in faeces of 
pigs.  

In terms of prevalence, C. coli were more predominant 
than C. jejuni in all farms. Farm Z had a higher incidence 
of C. jejuni and C. coli than the rest farms. This report is 
in line with previous reports which revealed that swine 



 
 
 

 

were predominantly contaminated by C. coli, whereas, C. 
jejuni were more frequently isolated from poultry (Aarestrup 

et al., 1997; Weijens et al., 1999; Van Looveren et al., 
2001; Pezzotti et al., 2003). The reason for this dis-
crepancy in the distribution pattern of C. jejuni and C. coli 
in swine has not been elucidated. However, it is not 
implausible that host adaptation or differences in rearing 
practices may be significant factors.  
Quite a large population of Campylobacter isolates from 
pig faeces where β-haemolytic on 5% sheep redblood 
cells. This observation is in line with the report of Samie 
et al. (2007). This is a confirmation that the 
Campylobacter strains could be pathogens of medical 
significance. The predominance of C. jejuni in human 
infections has led to the disregard of the impact that less 
prevalence species like C. coli can play in human 
infections. Nevertheless, a recent survey by Tam et al. 
(2003) showed that the morbidity resulting from C. coli 
has actually been underestimated.  

The PCR micrographs of the DNA from Campylobacter 
strains from all animals were as shown in Figure 1. The 
purified DNA from the Campylobacter strains amplified at 
the 1004 bp, which is the specific region for the conserved 
16S rRNA for members of the genus Campylobacter. The 
bands formed are in line with those of Marshall et al. 
(1999).  

After exposure to twelve (12) antibiotics, several strains 
showed multiple resistance to most of the antibiotics 
used. The resistance patterns displayed by Campylobacter 
isolates from all farms to fluoroquinolone (ciprofloxacin) 
and macrolides (erythromycin) classified as second line 
and first line antimicrobials are of particular importance, 
since patients suffering from campylobacteriosis are 
usually treated with these antimicrobials agents.  

In the study, 22.2, 40 and 31.1% of C. coli were 
resistant to ciprofloxacin, while 11.1, 60 and 50% of C. 
coli were resistant to erythromycin in farm X, Y and Z, 
respectively. For ampicillin, 55.6, 40 and 100% of C. coli 
were resistant to this antimicrobial in farm X, Y and Z, 
respectively. Norma et al. (2007) in their study in Canada 
observed a high incidence of C. coli resistant to 
ciprofloxacin and erythromycin. The resistance of C. coli 
to ampicillin as exemplified in this study was higher than 
those reported from France and Denmark (20 and 17%), 
respectively but lower than that reported from Spain 
(65.9%) for farm X, Y but not for farm Z (100%). 
Resistance to β-lactam antimicrobials in pathogenic 
bacteria develops through bacterial conjugation. This 
resistance-transfer mechanism is very crucial because it 
permits genetic exchange of information between species 
of bacteria (Davis and Conner, 1994; EFSA, 2007; Norma 
et al., 2007). In South Africa, data on anti-microbial 
susceptibility of Campylobacter isolated from pigs are 
scanty. Due to high prevalence of resistance shown by 
Campylobacter sp. to β-lactam antimicrobials (ampicillin) 
it is no longer recommended for use in farms (Davis and 
Conner, 1994). 

 
 

  
 
 

 

In this study, we observed resistance frequently to 
antimicrobials most commonly used in swine industry, 
tetracycline and erythromycin. Different resistance patterns 

were observed for one aminoglycoside antibiotic for all 
isolates tested for C. coli. 77.8, 52 and 50% were 
resistant to gentamycin in farms X, Y and Z, respectively. 
However, the report of Norma et al. (2007) in Canada 
showed a low level of resistance (0.2%) of Campylo-
bacter coli to gentamycin. The recommendation of 
gentamycin as an alternative therapeutic antimicrobial 
against human campylobacteriosis by Fernandez et al. 
(1994) in Chile should be considered with caution judging 
from the high prevalence of resistance to gentamycin 
observed in this study. Sato et al. (2004) and Inglis et al. 
(2005) observed high levels of resistance to tetracycline 
(45%), which is in line with our observation of ≥ 50%.  

Saenz et al. (2000) in their study reported no erythro-
mycin resistance among C. jejuni isolates from broilers 
and high levels of resistance C. coli isolates from pigs 
(81.1%) compared to those from humans (34.5%). 
However, in our study, one C. jejuni isolate out of two 
was resistant to erythromycin in farm X; all three C. jejuni 
isolates in farm Y were resistant to erythromycin, while 
one of the five C. jejuni isolate in farm Z was resistant to 
erythromycin. In addition, more resistance was associated 

with C. coli than with C. jejuni. A study in Quebec found 
that 61% of C. coli isolated from pigs but none of C. coli 
isolates from chickens were resistant to erythromycin 
(Guevrement et al., 2006). Resistance of Campylobacter 
species to antibiotics may suggest the widespread use of 
antibiotics as growth promoters in farm animals (Piddock 
et al., 2003; Pezzotti et al., 2003; Moore et al., 2001).  

Resistance profiles to tetracycline and fluoroquinolones 
were very high in our study and this is consistent with 
other reports (Saenz et al., 2000; Norma et al., 2007). 
This may have stemmed from the fact that these 
antibiotics have been used in the swine industry for 
several years.  

All the Campylobacter isolates were susceptible to 
nalidixic acid in this study, so that cross-resistance 
between nalidixic acid and ciprofloxacin was not found in 
all the quinolone susceptible Campylobacter strains. This 
finding is however in contrast to the observation of Saenz 
et al. (2000) who found cross-resistance between 
ciprofloxacin and nalidixic acid resistant strains in Spain.  

Moore et al. (2005) mentioned the use of mobile genetic 

elements in campylobacters as mechanism for the extrusion 

of antibiotics out of the bacterial cell. Considering the 
feeding habits of pigs, it is plausible that they can ingest 
commensals carrying resistant genes from the feeds, 
facilitating transfer to pathogenic bacteria in the lumen of 
farm animals. Such pathogenic acquisition of resistant 
genes from exogenous source can result in global 
prevalence of resistant pathogens. Angulo et al. (2004) 
and Blake et al. (2003) made similar arguments regarding 
acquisition of exogenous genes by lumen pathogens in 
farm animals. 



 
 
 

 

Surveillance and monitoring of antimicrobial use must be 
done to ensure the controlled use of antibiotics. The use 
of antibiotics as growth promoters and prophylaxis for 
animals should be carefully evaluated and monitored 
because acquisition of antibiotic resistant strains of 
campylobacters by man through ingestion of animal’s 
food have serious health implications. .  

Finally, the high prevalence of antibiotic-resistance among 

isolates of campylobacters in pig in the Venda region of 
South Africa needs further investigation. This study will 
be expanded to include the restriction analysis, antibiotic 
resistant genes and phylogeny of the Campylobacter 
isolates. 
 

 
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